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  • Title: Vesicular glutamate filling and AMPA receptor occupancy at the calyx of Held synapse of immature rats.
    Author: Yamashita T, Kanda T, Eguchi K, Takahashi T.
    Journal: J Physiol; 2009 May 15; 587(Pt 10):2327-39. PubMed ID: 19332485.
    Abstract:
    At central glutamatergic synapses, neurotransmitter often saturates postsynaptic AMPA receptors (AMPARs), thereby restricting the dynamic range of synaptic efficacy. Here, using simultaneous pre- and postsynaptic whole-cell recordings, at the calyx of Held synapse of immature rats, we have investigated the mechanism by which transmitter glutamate saturates postsynaptic AMPARs. When we loaded L-glutamate (1-100 mM) into presynaptic terminals, the quantal EPSC (qEPSC) amplitude changed in a concentration-dependent manner. At physiological temperature (36-37 degrees C), the qEPSC amplitude increased when intraterminal L-glutamate concentration was elevated from 1 mM to 10 mM, but it reached a plateau at 10 mM. This plateau persisted after bath-application of the low affinity AMPAR antagonist kynurenate, suggesting that it was caused by saturation of vesicular filling with glutamate rather than by saturation of postsynaptic AMPARs. In contrast to qEPSCs, action potential-evoked EPSCs remained unchanged by increasing intraterminal L-glutamate from 1 mM to 100 mM , even at room temperature, indicating that multi-quantal glutamate saturated postsynaptic AMPARs. This saturation could be relieved by blocking AMPAR desensitization using cyclothiazide (100 microM). The concentration of ambient glutamate in the slice, estimated from NMDA receptor current fluctuations, was 55 nM; this was far below the concentration required for AMPAR desensitization. We conclude that rapid AMPAR desensitization, caused by glutamate released from multiple vesicles during synaptic transmission, underlies postsynaptic AMPAR saturation at this immature calyceal synapse before the onset of hearing.
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